Whitefly has a broad host range of more than 600 species, and 40 biotypes have been reported worldwide. Among the various ecological types, B and Q are the most problematic. Especially, Biotype Q, which causes serious damage in the green house, mediates more than 40 kinds of viruses, including Tomato yellow leaf cull virus (TYLCV). The development of resistance to whitefly, adverse effects on non-target organisms, and strengthened legal regulations are making chemical control difficult in application. For these reasons, the use of alternative strategies is needed to suppress the Biotype Q whitefly population. Entomopathogenic fungi were investigated and developed to control hard-control insect pest, such as whitefly. These fungi are natural pathogen of their invertebrate host and contribute to the regulation of their host population in the environment. In addition, these agents are typically harmless to humans and non-targeted organisms and do not adversely impact the environment. Therefore, in this study, we screened pathogenic fungi against whitefly using our entomopathogenic fungal library, and virulence tests were conducted with conoidal suspension (1×107conidia/ml) in laboratory conditions. As a result, 10 isolates showed 80-100% mortality at 5 days after treatment. We will analyze the characteristics of these fungal isolates and evaluate their potential as insecticides against whitefly. The selected isolates against the high mortality of whiteflies can be effectively used in integrated pest management programs for whiteflies.

Japanese pine sawyer beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae) is considered as a serious pest in pine trees. To develop an eco-friendly strategy to manage this forest insect, we collected entomopathogenic fungi from Korean soil and assessed their virulence against the adults of the insect in laboratory conditions. As a result, two isolates with conidial suspension (1.0×107conidia/ml), showed 87% and 90% mortality 12 days after fungal treatment, respectively. We assessed the potential of the fungi-derived destruxin and protease as additives to the fungal isolates, and they showed insecticidal activity via feeding and spraying treatments. Finally, we produced fungal conidia in massive solid cultures and formulated wettable powders, and now studying optimal conditions of oil-based formulation with two isolates. Based on these results, we are evaluating the control efficacy of the fungal agents against M. alternatus in field conditions.

The composition of the deicer sprayed on the highway is spreading over the highway by the scattering or snow removal activity, or car movement and consequently affecting the vegetation environment around the highway. These are the cause of the damage of fruit trees and crops, and also the cause of corrosion of highway structures. The goal of this study is to estimate the detected range of deicer components from a highway. The concentration of the deicer components in gauze and soil were investigated according to the crossing distance from the highway. The data collected were then used to estimate the concentration range of deicer components in a rage of distance up to 100 m from the highway where the deicer was spread. The sample time and weight of gauze were measured before and after installation, and the soil was collected at more than three points in parallel with the highway at the gauze installation point. The components in gauze and soil were investigated in addition to the deicer components (Ca2+, Na+, Cl-) as well as Mg2+ and K+. As Ca2+ and Cl- components of deicer were affected by agricultural use, Na+ component was selected as a tracer and further SAR (Sodium Adsorption Ratio) of soil was analyzed to examine the degree of influence on vegetation indirectly. The gauze concentration was evaluated by the concentration of the deicer ingredient at the background concentration of the blank gauze. The total amount of the deicer sprayed in the study road for 4 months (winter season) was about 93 ton/km. In the gauze test, the spread of the deicer was detected at a distance of 100 m in study area, but the concentration of the deicer in the gauze by distance decreased rapidly within 10 m from the highway. And concentration of the deicer components in gauze and soil came down after rainy season (August ~ Sep.). The results showed that the components of the deicer could be spread widely by the wind. The effective range of the deicer on vegetation based on SAR in soil was estimated to be less than 20 m from the highway. This study examines the concentration changes of the deicer components in gauze and soils and shows that deicer components sprayed on the highway are accumulated and moved over time by wind, snow removal, terrain, water system and land use around the highway.

최근 봄철 기온이 높아짐에 따라 마늘 뿌리응애 피해가 늘어나고 있는 실정이다. 이러한 뿌리응애 피해를 최소하 하고자 발생양상과 22종의 약제에 대하여 살비효과를 조사하였다. 마늘 재배시기 중 뿌리응애의 발생은 충북 보은 등 5지역에서 2015년부터 2017년까지 3년간 3월부터 6월까지 뿌리응애 발생밀도를 조사하여 발생양상을 확인하였다. 뿌리응애는 2016년에 가장 많이 발생하였으며, 발생시기는 월동 후 3월 상순부터 발생하여 6월 중순경까지 지속적으로 밀도가 높아지는 양상을 나타내었다. 조사된 5지역 중 가장 높은 밀도로 조사된 보은지역은 6월 상순경 주당 13.3마리가 발생하였다. 뿌리응애에 대하여 우수한 살비효과를 나타낸 약제는 분무처리시 약제처리 10일 후에서 dimethoate, azocyclotin, abamectin, hexythiazox, cabosulfan, spirotetramat, benfuracarb가 90% 이상의 살비효과를 나타내었다

Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae) in forest is considered as a serious forest pest in pine trees. In this study, we collected entomopathogenic fungi from soil and assessed their virulence against the adults of the inset using a spray method in laboratory condition. Two entomopathogeinc fungi isolates showed high virulence against the adults. In the concentration of 1.0 × 107conidia/ml, the mortality rate of the adults were 53% and 60%. In semi-field condition, one isolate showed a virulence of 60% against the adult. Consequently, we confirmed the possibility of the fungal isolates in controlling the beetles. In near the future, we will investigate several factors which is possibly related to the control of the forest insect pest using entomopathogenic fungi in field conditions, given the importance of fungal formulation and practical application methods.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopenia syndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest. Therefore, this study provides a screening method to select entomopathogenic fungi, having high virulence against H. longicornis. The virulence test was conducted by dipping method with a conidial suspension (1×107 conidia/ml). As a result, cadavers of H. longicornis infected with entomopathogenic fungi were obtained by this method. Based on this results, we selected several isolates having high virulence to H. longicornis. We suggest that, selected isolates in this study can be used for the control of H. longicornis.

Entomopathogenic fungi have been known as promising candidates for biological control of insect pests. Recently, researchers consider the fungal thermotolerance in formulations and field applications. In this study, we investigated the production of thermotolerant Isaria javanica and I.fumosorosea conidia through grain-based solid cultures and exposure to light stress. As results, of the ten grain substrates, Italian millet, rice, perilla seed and sesame, rice, sorghum produced highly thermotolerant conidia in the strains. The two strains were exposed to a light stress and a heat stress. And they showed enhanced thermal stability compared to control, when exposed to 45°C for 2 hours. This work suggests that heat-resistant entomopathogenic fungal conidia can be produced by grain-based solid cultures and exposure to light stress.

Melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae) is a serious insect pest of various crops in the world. The management of this pest mainly have relied on chemical agents. However, the overuse of chemical agents for long times causes insect resistance, and negative effects on environment. Therefore, alternative control methods, such as biological control, that are less harmful to the environment and have different mode of action are needed to control this pest. In this study, we isolated entomopathogenic fungi from Korean soil, and characterized them via morphological and molecular identifications and pathogenicity check against Tenebrio molitor larvae. The isolated fungi were subjected to virulence assay against T. Palmi with a conidial suspension of 1×107 conidia/ml in laboratory conditions. This entomopathogenic fungi library would be a good resource to select effective strains for the control of T. palmi in fields.

Entomopathogenic fungi play an important natural role in regulating their insect host populations, and their ecology was also associated with plant and soil. These microorganisms have been living by reacting to insect, plant and environmental factor. The advanced bioinformatics technology such as next-generation sequencing and RNA sequencing has revolutionized in understanding of entomopathogenic fungi. Recently studies provided a lot of information on evolutionary relationships and virulence-related characteristics. We are starting to know where these microorganisms from is, and how they live in nature. The bioinformatics technology will give us further our understanding of the natural roles of these fungi in nature.

The entomopathogneic fungal colonization has become an interesting issue in ecological pest management. Plant-orientedfungi induce plant growth promotion and act as natural control agents against insect pest. Herein this work, we evaluatedthe ability of fungal colonization on bark, branches and leaves of pine tree (Pinus densiflora) to select optimal candidatesfor controlling Japanese pine sawyer (Monochamus alternatus), which is the pine wilt disease vector in Korea. As a result,some Beauveria and Metarhizium isolates could colonize on the substrates of P. densiflora and showed pathogenicity againstM. alternatus. This result suggests that the selected entomopathogenic fungi are possibly candidates for long-term managementof M. alternatus in pine tree forest.

Japanese pine sawyer, Monochamus alternatus HOPE (Coleoptera: Cerambycidae) are considered as important pine forestinsect pests in several countries which transmits the pine wood nematode, Bursaphelenchus xylophylus. In this study, wetried to control Japanese pine sawyer using entomopathogenic fungi to prevent the spread of pine wilt disease in Korea.Several entomopathogenic fungi were exposed to the adults of Japanese pine sawyer and mycosis was observed in themembranous cuticles after 6 days. In particular, two isolates showed high virulence against the adults. The selected 2isolates were sprayed to the adults at 1×105, 1×106 and 1×107 conidia/ml. In the concentration of 1×107 conidia/ml, thesurvival rates of insects were 40% and 47%, respectively. Based on these results, the possibility of controlling the Japanesepine sawyer was confirmed.

Chemical pesticides have been used for persimmon pest control, however the overuse caused insect resistance andenvironmental toxicity. Herein this work, we investigated the biological potential of eco-friendly organic pesticides (biologicals)and entomopathogenic fungi in controlling persimmon pests, Stathmopoda masinisaa and Riptortus pedetris. A botanicalmixture of Chinese scholar tree, goosefoot and subtripinnata extract, Bacillus thuringesis NT0423 and one isolate of Beauveriabassiana showed high insecticidal activity against the pests in laboratory conditions. The selected botanical extract andmicroorganism were combined with a chemical pesticide (buprofezin+dinotefuran WP) and were assessed their controlefficacy in persimmon fields. Compared to the three times of chemical spray, the combination of more than one timeof chemical spray and the selected biologicals controlled the persimmon pests with an acceptable levels. Consequentlyone time of biological spray can be integrated to the spray program where only chemicals are strongly considered.

The hard tick, Haemaphysalis longicornis (Ixodida: Ixodidae) is one of the vectors of severe fever with thrombocytopeniasyndrome virus (SFTSV) causing in humans, however little consideration was given to the biological control of the pest.We evaluated three bioassay methods, such as spray, dipping and dropping. As a result, the dipping method was themost efficient way to measure the fungal virulence. Finally, we constructed a Haemaphysalis longicornis pathogenic fungallibrary to further facilitate the resources to be used as potential biological control agents. A conidial suspension of 1×107conidia/ml was exposed to the ticks by dipping. We confirmed that some isolates were highly virulent to the ticks. Thisresults indicated that some fungi could be used to effectively control the Haemaphysalis longicornis.

The mesenchymal stem cells (MSC) has been investigated as a source of stem cell therapy to replace and treat damaged cells. Human endometrial epithelial and stromal cells was isolated from hysterectomy tissue and the direct evidence of stem/progenitor cells in the human endometrium was identified. Endometrium derived stem cells (EnMSCs) are known to have a high proliferative ability, genetic stability, lack of tumorigenicity and low immnunogenicity during long-term cultivation. Here, we aimed to identify MSC in canine endometrium and characterize its potential to differentiate into decidua cells. EnMSCs were isolated from thrown-away spayed uterus of adult canine depending on their estrus cycle, and identified by flow cytometry, immunocytochemistry and flow cytometry with MSC specific markers. We then characterized the ability of EnMSCs by the doubling-time analysis, colony-forming units and MSC differentiation assays. Isolated EnMSCs expressed stem cell specific genes (Sox2, Oct4, Nanog, MCAM, Endoglin, Susd2 and IGTB) and MSC surface markers (CD90, CD44 and CD117). EnMSCs are also differentiated into adipogenic, osteogenic and chondrogenic cells morphologically under modified conditions with the expression of lineage specific genetic markers. EnMSCs showed higher proliferation ability than canine amniotic fluid derived MSCs which were used as a positive control. EnMSCs were cultured at low density (10, 20, cells/cm2) and initiated to form small colonies of loosely-arranged cells and gradually formed large colonies of densely-packed cells which underwent self-renewal with high proliferative potential which is similar to the clonogenicity feature of human endometrium-derived stem cells. EnMSCs were then induced to differentiate into decidua cells with 0.5 mM dbcAMP. After 14 days, EnMSCs changed their morphology into the elongated and rounded shape. The induced decidual cells expressed PRL and IGFBP1 which are typically expressed in decidua cells. In conclusion, we successfully isolated and characterized MSC in the canine endometrium which differentiated into decidua cells. These results showed that endometrium may be a promising source of stem cells, and furthermore raise the possibility of canine EnMSCs as a novel hypothetical decidualisation model of infertility associated with decidualisation insufficiency and implantation failure.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

To obtain information on the sanitary of indoor environment in greenhouses used for shiitake cultivation, bacteria associated with larvae and imagoes of Sciaridae flies, pest to shiitake mushroom, were isolated and identified. A total of 1,048 bacterial colonies were isolated from the flies’ larvae and 984 bacterial colonies were isolated from the flies’ imagoes. Based on molecular analysis of the 16S rDNA sequence, Achromobacter xylosoxidans and Tetrathiobacter kashmirensis of β-proteobacteria, Enterobacter asburiae and Raoultella ornithinolytica of γ -proteobacteria, Curtobacterium sp. and Microbacterium thalassium of Actinobacteria, and Penibacillus taichungensis of Firmicutes were identified from the colonies of the flies’ larvae. While, Bacillus megaterium, B. thuringiensis, Lysinbacillus sphaericus and L. fusifomis of Firmicutes, Microbacterium thalassium and Citricoccus parietis of Actinobacteria, and Enterobacter asburiae of γ-proteobacteria were identified from the flies’ imagoes. Some of the isolated bacterial species were known be human pathogens. Overall, the results of this study suggested that mushroom fly carrying human pathogenic bacteria is one of sources impact on the sanitary of indoor environment of greenhouses used for shiitake cultivation.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), was originally observed in a few Asian countries, but is now found even in North America and Europe. Genetic information on geographic variation and relationship may broaden our understanding of origin and migration. As a first step, in this study, a portion of mitochondrial COI gene was sequenced to understand genetic relationship and diversity in Korea. Sequencing of 104 individuals provided 57 haplotypes, with the maximum sequence divergence of 1.5%, suggesting high haplotype diversity and moderate sequence divergence. Comparison to GenBankregistered D. suzukii haplotypes (possibly from Spain, Portugal, USA) has shown 100% sequence identity to most of the haplotypes found in this study, but two USA sequences were found to be independent haplotypes, with the sequence divergence ranging from 0.5% ~ 1.4% from our samples in the 553-bp comparison. Phylogenetically, no separable group was found, but, population genetically, the only Chinese population, Sandong, was significantly differentiated (p < 0.05) from all Korean populations, without sharing any haplotype. Among 28 pairwise comparisons of Korean populations only two comparisons showed a significant genetic differentiation, indicating that no population in Korea is completely isolated. Geographically, one haplotype (SWDBA06) was relatively widespread (five among nine localities) and a few haplotypes were found in more than one locality, but most haplotypes were restricted in a locality as a single individual. Overall, high rate of per generation female migration (Nm = 0.75 ~ infinite) and low level of geographic separation (FST=0~0.40) among localities were characteristic. Current data is limited mainly to Korean localities, thus, an expanded study may provide further scrutinized analysis for the fly.

The anti-proliferative efficacy of t,t-conjugated linoleic acid (t,t-CLA), c9,t11-CLA, and t10,c12-CLA was compared in several human cancer cell lines. Gastric NCI-N87, liver Hep3B, pancreas Capan-2, and lung NCI-H522 cancer cells were incubated with 50 μM CLA isomers over a period of 6 days. The t,t-CLA inhibited the growth of all cancer cell lines to different extents, but c9,t11-CLA and t10,c12-CLA inhibited or stimulated the growth of the cancer cell lines. NCI-N87 cells were the most sensitive to growth inhibition and apoptosis from all CLA isomers tested. In NCI-N87 cells, CLA isomers reduced the release of arachidonic acid (AA) via the inhibition of cytosolic phospholipase A2 (cPLA2 ) activity, consequently reducing the production of PGE2 through the inhibition of cyclooxygenase-2 (COX-2). The efficacies of CLA isomers were in the following order (from most to least effective): t,t-CLA, t10,c12-CLA and c9,t11-CLA. Overall, these results imply that the anti-proliferative efficacy of t,t-CLA on cancer cells, especially NCI-N87 cells, was greater than other CLA isomers due to its induction of apoptosis through the inhibition of cPLA2 and COX-2 activities.

Escape pattern of the ambrosia beetle Platypus koryoensis (Murayama) based on individual gallery was investigated. The survey was conducted in four oak forests located in Chuncheon, Hanam, Paju and Seongnam, and escape patterns from individual gallery were surveyed using emergence trap attached at the entrances of galleries formed in 2010. The number of adults caught in each trap was periodically counted from May 2011 to September 2012. The adults ambrosia beetle was caught over two years after infestation. The escape patterns in the first and second year after infestation showed different types; in the first year, escape of adults from gallery was observed one or two times whereas escape from gallery was observed two times in the second year with variation in aspect of escape density and timing. Moreover, variations in escape pattern between trees or between galleries in the same tree was observed. Colonization success rate in living tree was higher than that of dead tree, but the number of progeny per gallery in living tree was lower than that of dead tree. Colonization success rate in the galleries of second year was lower than that of first year, but the number of progeny per gallery of second year was higher than that of first year.